I am currently planning some LUMA global methylation analysis using a Qiagen Pyromark Q24 system. Can anyone give advice on which program to use to generate the assay-I know it should be the AQ assay but what sequence is best to use to generate the dispensation order? Upon running the reactions, is there a tool for calling the peak heights?
Dear Alan, The Qiagen Pyromark Q24 system have one integrated PyroMark Q24 Advanced Software. As part of the package, it includes software for the amounts and order of the master mix dispensing. The program is installed on a computer, you generate dispensing program on a USB memory and insert it into the machine, it reads and runs the test and generates values, to which you apply the formulas set them on a sheet of excel to calculate the values of global DNA methylation. There are several dispensing programs reported in papers and each laboratory used that works best, there is not one
Dear Alan,
I think you can use the Qiagen PyroMark Assay Design software to design the CpG assay. It will give you a set of primers (forward, reverse and sequencing primers) targeting the CpG sites that you want to examine. During the assay designing, you will find the biosulfite converted sequence information which can be copied to the PyroMark Q24 2.0.6 software for generation of the dispensation order.
Hope the information above is helpful.
Thanks for the input. I am very familiar with running pyroseq CpG assays on the Q24 machine, I was hoping to find out if anyone had any info on running LUMA using this equipment.
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